LOCUS       V047100                 8694 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V047100
VERSION     V047100
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        50..429
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        430..633
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        678..696
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             756..3374
                     /label="Zhx1(NM_001042438)"
                     /note="Zhx1(NM_001042438)"
                     /gene="Zhx1"
     CDS             3381..3446
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3470..4058
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4124..4172
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4374..4802
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4816..5145
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5212..6006
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /note="confers resistance to neomycin, kanamycin, and G418
                     (Geneticin®)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /transl_table=1
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
                     /label="NeoR/KanR"
     polyA_signal    6182..6303
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6352..6368)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6376..6392
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be
                     relieved by adding lactose or
                     isopropyl-β-D-thiogalactopyranoside (IPTG)."
                     /label="lac operator"
     promoter        complement(6400..6430)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6445..6466
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6754..7342)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7513..8373)
                     /codon_start=1
                     /gene="bla"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /product="β-lactamase"
                     /transl_table=1
                     /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY
                     SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR
                     WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS
                     ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA
                     SLIKHW*"
                     /label="AmpR"
     promoter        complement(8374..8478)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
       61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      601 cgtgtacggt gggaggtcta tataagcaga gctctctggc taactagaga acccactgct
      661 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgttta
      721 aacttaagct tggtaccgag ctcgaattcg ccaccatggc aagcagacga aaatcaacaa
      781 caccttgcat ggtccttgcc agtgagcagg atccagatct tgagctgata tcggatttgg
      841 atgaaggccc tcccatcctt acacctgtag agaatgctaa agcagagagt gtctccagcg
      901 atgaagaagt tcacggatct gtggattctg acaatcagca aaataagaaa gtggaagggg
      961 gctatgaatg taaatattgt acttttcaaa ctccagatct aaatatgttt actttccatg
     1021 tagattcaga acatcccaat gttgtgctga attcctccta tgtttgtgtt gaatgcaatt
     1081 ttcttaccaa aaggtatgat gcactttctg agcataatct gaaataccac ccaggagaag
     1141 aaaatttcaa gctgactatg gtgaagcgga ataaccagac aatctttgaa cagacaataa
     1201 atgatctaac ttttgatggc agttttgtta aagaggagaa tacagaacaa ggagaatcca
     1261 tagatgtttc ttcttcggga atctccatta gtaaaactcc catcatgaaa atgatgaaaa
     1321 ataaggtaga gaacaaacgg attacagtcc accataactc agctgagggc acttctgaag
     1381 agaaagagaa tggagtaaaa gcaagccagg aagaaaatgc agaaagtgtg agttcttcag
     1441 ctttagaatc caatacaagt acttccacca tcaacagagt gcatcccagt cctgccagca
     1501 cagtggtgac tcccacagct gttcttcctg ggttagcaca ggtgataact gcggtgtctg
     1561 ctcaacagaa ctcaaacttg ctccctaaag tcttaatccc cgttaatagc attcctacct
     1621 acaatgctgc attggataac aatcctcttc tacttaatac ctacaacaaa ttcccttatc
     1681 ctacaatgtc agaaattaca gttctttctg ctcaagcaaa atatacagag gaacaaatca
     1741 agatctggtt ttcagcccaa cgtttaaagc atggtgttag ttggactcct gaagaagtag
     1801 aggaggcgag aagaaaacaa ttcaatggaa cagtacatac tgtacctcag accataactg
     1861 ttattcctac acacatttcc acagggagta acgggttacc atctatctta cagacatgcc
     1921 aaattgttgg tcagccaggt ctggtcctta ctcaagtagc tggaacaaac accttgccag
     1981 taacagcacc cattgccctg acagtggcag gggttccaaa tcaaacaaat gtacaaaaga
     2041 gccaagtgcc tgctgctcag cctgctacag acacaaaacc agccacagca gcagttccat
     2101 cctcgccaag tgtcagacct gaagctgcac tagtgaaccc agattcattt ggtattcggg
     2161 ccaaaaagac taaagagcag ctggcagaac tgaaagttag ctacctaaaa aatcagtttc
     2221 cccatgactc agagatcatc aggcttatga aaataacagg ccttaccaaa ggcgagatta
     2281 aaaaatggtt tagtgataca aggtacaacc agagaaattc aaagagtaac cagtgcttac
     2341 atctcaacaa tgactcctct gccactatca tcatagactc cagtgatgaa accccagaac
     2401 ccccagcggc agctgcttca cagcagaaac agtcctggaa tccctttcct gactttgctc
     2461 cccagaagtt taaagagaaa actgcagagc aactccgtgc ccttcaggca agttttctca
     2521 acagttctgt acttactgat gaagaagtaa ataggttaag agcgcaaacc aaacttacta
     2581 gaagagaaat tgatgcttgg tttacagaga aaaataaaac aaaagcttta aaggatgaga
     2641 aaatagaagt agatgaaagt aatgtaggta gttccaaaga agagcctgga gaaagttctc
     2701 ctggagatga aacagttgca cctaagtcag gagggacagg caagatatgt aagaaaacac
     2761 cagagcagct gcacatgctt aaaagtgcat ttgtccgaac acagtggcca tcagcagaag
     2821 agtatgacaa gctggctgaa gagagtggcc ttgccagaac agatatagtt agttggtttg
     2881 gggatacccg ttatgcttgg aagaatggaa acttgaaatg gtattactac tatcaaagct
     2941 ccaattcaag tagtttgaat ggtctatctt cccttaggag aagagggaga ggcagaccca
     3001 aaggaagggg caggggcaga ccacgtggtc ggcccagagg tggaaaaaga atgaacacct
     3061 gggacagggt accatcgctt ataaagttta aaactggaac agcaatactt aaggattatt
     3121 acctgaagca caaatttctt aatgagcaag accttgatga acttgtcaac agatcacaca
     3181 tgggttatga gcaggtcaga gagtggtttg cagaaagaca gagaagatct gagttaggta
     3241 tagaattatt tgaggaaaat gaggaggaag atgaagttgt tgatgatcag gaagaagatg
     3301 aagaagaaac agatgatagt gacacttggg aacccccacg acatgtgaag cggaagcttt
     3361 ctaaatcgga tgacggatcc gactacaaag accatgacgg tgattataaa gatcatgaca
     3421 tcgactacaa ggatgacgat gacaagtagt gagggcccga tatctcgaca atcaacctct
     3481 ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct
     3541 atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat
     3601 tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt
     3661 caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat
     3721 tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc
     3781 ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga
     3841 caattccgtg gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg cctgtgttgc
     3901 cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga
     3961 ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc
     4021 tcagacgagt cggatctccc tttgggccgc ctccccgcct ggaaacgggg gaggctaact
     4081 gaaacacgga aggagacaat accggaagga acccgcgcta tgacggcaat aaaaagacag
     4141 aataaaacgc acgggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg
     4201 gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt
     4261 ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc
     4321 ggcaggccct gccatagcag atctgcgcag ctggggctct agggggtatc cccacgcgcc
     4381 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     4441 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     4501 cggctttccc cgtcaagctc taaatcgggg catcccttta gggttccgat ttagtgcttt
     4561 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     4621 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     4681 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     4741 tttggggatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     4801 ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     4861 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     4921 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     4981 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     5041 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc
     5101 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct
     5161 tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa
     5221 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     5281 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     5341 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag
     5401 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     5461 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     5521 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     5581 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     5641 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     5701 gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat
     5761 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     5821 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     5881 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     5941 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     6001 ttctgagcgg gactctgggg ttcgcgaaat gaccgaccaa gcgacgccca acctgccatc
     6061 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
     6121 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
     6181 caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     6241 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     6301 ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct
     6361 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     6421 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     6481 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     6541 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     6601 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     6661 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     6721 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     6781 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     6841 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     6901 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcaat gctcacgctg
     6961 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     7021 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     7081 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     7141 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt
     7201 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     7261 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     7321 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     7381 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     7441 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     7501 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     7561 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     7621 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     7681 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     7741 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     7801 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     7861 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     7921 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     7981 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     8041 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     8101 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     8161 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     8221 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     8281 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     8341 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     8401 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     8461 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg
     8521 agatctcccg atcccctatg gtcgactctc agtacaatct gctctgatgc cgcatagtta
     8581 agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt
     8641 taagctacaa caaggcaagg cttgaccgac aattgcatga agaatctgct tagg
//
